glycomics fnl 2010july 1280x720vid fnl - meetupfiles.meetup.com/3413352/glycobiology class slides...

Introduction to the Science of Glycobiology

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Glyconutrients

‐‐ ANGIEANGIE LAW LAW ‐‐

Glyconutrients

WWW.ANG I E SOPT I ON . COMWWW.ANG I E SOPT I ON . COM

l lGlucose(from table sugar)

Typically, only these twoare found in our modern diet.

Mannose N‐Acetyl glucosamine

Xylose N‐Acetyl galactosamine

Galactose(from milk products)

Fucose N‐Acetyl neuraminic acid


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www.Proevity.com

Continuing Medical Education site

G d M f t i P ti (GMP) Good Manufacturing Practices (GMP)

NSF International Certification

P Patents U.S.

International


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MIT’s Technology ReviewgyFebruary 2003

GlycomicsGlycomics one of the one of the top 10 technologies that top 10 technologies that top 10 technologies that top 10 technologies that will change the worldwill change the world

The Study ofBiological Sugar ChainsBiological Sugar Chains

‐‐ LARRY A. LAW LARRY A. LAW ‐‐

I N FORMAT IONAL SOURCE : ROBERT K . MURRAY, MD, PHDIN FORMAT IONAL SOURCE : ROBERT K . MURRAY, MD, PHD


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The only difference Red Blood Celldifference between

Type A (GalNAc)

and

Type B (Gal)

GalNAc

GlcNAc

Red Blood Cell

OO

AA

ABABType B (Gal)

is one sugar, but it can make a life/death difference in a transfusion

Fucose

Galactose

BB

Glyco—Sweety

Glycoprotein—Protein molecule with one or more sugars attached. Found in all cells, tissues, and body fluids. 50% of all human proteins have sugars attachedsugars attached.

Glycosylation—Attachment of sugars to proteins and lipids via enzymatic attachment.


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It’s not what we don’t know that It’s not what we don’t know that prevents us from succeeding; it’s prevents us from succeeding; it’s what we KNOW that just what we KNOW that just ain’tain’t so.so.jj

——Mark TwainMark Twain

Old: Sugar only used for energy productionN Th ll ili 8 i l l l i New: The cell utilizes 8 essential sugars to glycosylate proteins and lipids within the Golgi Apparatus and Endoplasmic Reticulum.

These glycoforms cover the cell membrane and interact with other cells for specific biologic functions including:

Cell to cell communication and identification Immune system function Cell/tissue/organ repair and function


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Glycoprotein Cell Receptors are surface sugars that serve as points of

BacteriumVirus

White Blood Cell

Abnormal Cell

attachments for cells, bacteria, viruses, and many other molecules.

ToxinGlycoprotein

Normal“Hairy” Cell

Old: Glucose only pathway—all sugar changed to glucose in the bodybody

New: Study in France demonstrated radioactive galactose and mannose were directly absorbed into the body without being broken down into glucose. Scientists conclude that specific dietary sugars could represent a new class of nutrients (Dec 1998).

Old Stomach enzymes cannot break down beta bonds holding Old: Stomach enzymes cannot break down beta bonds holding sugar chains together

New: Bacteria in the intestinal tract break these beta bonds so sugars can be absorbed directly by the body (Jan 2007)


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Old: Proteins serve as the communication signaling l l h ll l l d dmolecules at the cell level. But 20 amino acids do not

provide enough different chemical combinations to support a vocabulary large enough to account for cellular communication.

New: The hexagonal, 3‐D structure of the 8 sugars d h l l l k b l dand their multiple linking possibilities provide a

potential of 1x1014 (100,000,000,000,000) chemically unique “letters” in this sugar code of biologic information.

Undermodulated Immune SystemUndermodulated Immune System Cancer

Overmodulated Immune SystemOvermodulated Immune System Multiple Sclerosis (MS)

Rheumatoid Arthritis (RA)


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RELEVANT FACTORSRELEVANT FACTORS

Six of eight NOT found in our modern diet

Cooked/Processed/Refined foods

Green harvesting

Lower nutritional value of fruits and vegetables (Food Pyramid & Soil Depletion)

Increased oxidative stress in modern world Antibiotics, drugs, alcohol Chemicals, plastics, pesticides, perfumes

Primary versus backup system


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Autoimmune disease exponential increasein last 30 years in last 30 years Chronic Fatigue Syndrome, Lupus, Fibromyalgia Diabetes 30 million to 246 million in last 3 decades Type I: 5‐10% of cases Type II: 90‐95% of cases (formerly adult onset)

1990 – 1998 Ages

Heart Disease Cancer

70% 30‐39

40% 40‐49

American medical system #1 in trauma carey

But World Health Organization ranks U.S. 72nd

in chronic disease U S spends in excess of $7 290/person on U.S. spends in excess of $7,290/person on health care (#2 Switzerland ‐ $4,417)

Britain spends $2,992 but Brits are healthier than Americans in everymajor health category


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WHAT DIFFERENCE DO THEY MAKE?WHAT DIFFERENCE DO THEY MAKE?

Cell to Cell Adhesion—h ll li d ti k t th

Structural Support—ll d th ti how cells align and stick together

Cell to Cell Communication—glycoprotein receptors and how hormones (eg. insulin) affect destination cells

collagen and the connective tissue between cells are glycoproteins

Lubricants—mucins Immunoglobulins Transport—

Fertilization Antibody Functions—

all antibodies are glycoproteins

plasma proteins (eg. Transferrin)

Some Enzymes, Hormones—Alkaline phoshatase, TSH, hCG


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Inflammation—Attachment of neutrophils and lymphocytes to walls of small blood vesselsof small blood vessels

Platelet Aggregation—Attachment of platelets to walls of blood vessels and each other. Hemostasis (clotting) and thrombosis (heart attacks and strokes)

Influenza—Attachment and release of virus AIDS—Attachment of HIV‐1 to cells Bacterial Infections—Urinary Tract Infections Peptic Ulcer Bacterial Infections Urinary Tract Infections, Peptic Ulcer Rheumatoid Arthritis—Altered glycosylation of IgG Cancer—Altered cell surface glycoproteins, metastasis Genetic diseases—Cystic Fibrosis, Muscular Dystrophy, congenital

disorders of glycosylation

Chapter 46 “Glycoproteins”(p 523‐544)

(27th Edition)

(p. 523 544) “About 200 monosaccharides are

found in nature, however, only eight are commonly found in the oligosaccharide chains of glycoproteins.” (p. 524)

1996 edition was first time a medical textbook devoted an entire chapter to the biological significance these 8 sugars have in cellular communication.


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“There are enzymes in human tissue that can make

(27th Edition)

ythese sugars from glucose. However, there is evidence that the other sugars may be beneficial in some circumstances when added to the diet.This has led to the development of glyconutrientsupplements containing either members of the supplements containing either members of the sugars listed in Table 46‐4 (excluding glucose) or precursors of them. The efficacy of such supplements is under study.” (p. 524)

Why doesn’t your doctor know about this?

(27th Edition)

y y Wasn’t in medical school text book until 1996, so if they graduated before 2002…

7,000 articles printed in medical literature every day. If they read 2 articles each day, at the end of the year, they’d be 35 centuries behind in their reading! (Dr Fouts)centuries behind in their reading! (Dr. Fouts)

Who funds continuing education credit seminars for doctors? Pharmaceutical companies. Who funds research studies? Not broccoli farmers!


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Multiple studies have documented breast fed babies have i b i i l h l h b i stronger immune systems, better intestinal health, brain

development, and cognitive abilities than formula fed babies

Human breast milk contains 5 of the 8 biologically active beta bonded sugars (critically different from cow or goat’s milk) Lactose (galactose/glucose), glucosamine,

sialic acid fucosesialic acid, fucose

Chains of beta‐bonded sugar structures were originally thought to be of no benefit to the body

1/3 of milk solids are these sugars (higher % than protein)

A LEAGUE OF THEIR OWNA LEAGUE OF THEIR OWN


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Extracted and stabilized from aloe era plant vera plant

Aloe mentioned in literature for over 5,000 years as having healing and curative properties

Al Aloe vera craze

What grandma knew but couldn’t explain

Photo: Aloe © Sinisa Botas | Dreamstime.com

Incorporated into glycoprotein receptors on ll f

BIOLOGICAL FUNCTION

cell surfaces.

Modulates the immune system primarily by activating macrophages. Macrophages phagocytize

bacteria and cellular debris bacteria and cellular debris associated with tissue inflammation. Stahl PD Am J Respir Cell Mol Biol.

1990 Apr;2:317‐318

Photo: Macrophage © Sebastian Kaulitzki | Dreamstime.com


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Mannose prevents bacterial infections by binding

BACTERIAL INFECTIONS

p y gcompetitively to bacterial lectins. Therefore, it occupies the sites that would otherwise bind to hostcell receptors.

By preventing bacterial adhesion, infection is prevented. Beuth J Int J Med MicrobiolViral Parasitol Infect Dis. 1994 Oct;281:324‐333

Free sugar molecules bind to

BACTERIAL INFECTIONS

Free sugar molecules bind to lectins preventing adhesion.


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Blocks Salmonella adherence to chicken intestine

ANIMAL STUDIES

in vitro Reduces incidence of Salmonella infections in vivo by 50‐100% when added to drinking water for chickens Oyoto BA Poult Sci. 1989;68: 1357‐1360

Oyoto BA Avian Dis. 1989 Jul’33:531‐534

Decreases E. coli bacteria in newborn mice when applied topically to maternal vaginas prior to delivery Cox F J Infect Dis. 1990 Oct; 162:978‐981

Abundant in seaweeds and sea urchins/starfish

Exhibits considerable anticoagulant and antithrombotic activity

Topical administration of L‐Fucose increases skin pthickness and density of collagen

Increases free radical scavenging


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Stimulates adult hematopoietic stem cell formation.

FUCODIANS AND STEM CELLS

p

Healthy adult volunteers ingested 3 grams per day of Undaria fucoidans for 12 days with a significant increase in stem cells seen in peripheral bl d blood smears. IrhimehMK et al Fucoidan and Cxck4 and hemopoietic progenitor stem cell

population, 2004 Nov. The Sydney Convention Centre North, Darling Harbour, Australian Stem Cell Centre

Important in cell‐to‐cell communication Widely distributed in body tissue Thyroid gland has NAG receptor indicating a role in

transport of thyro‐globulin within the gland ThibaultV Endocrinology. 1993 Jan;132:468‐476

Glucosamine is substrate for glycosaminoglycan (building block of damaged cartilage) Inhibits metabolic breakdown

Directly involved with repair of damaged cartilage Talent JM ClinTher. 1996 Nov; 18:1184‐1199


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Findings of Dr Demetriou of the University of California Irvine Center Findings of Dr. Demetriou of the University of California Irvine Center for Immunology published in the Journal of Biological Chemistry,16 Nov 2007

NAG suppresses the growth and function of abnormal T‐cells responsible for autoimmune attack in MS and Type 1 Diabetes

NAG prevented T‐cell hyperactivity that results in autoimmune attack on brain myelin and the insulin‐producing cells of the pancreas. Preventing this attack of the body on itself protects against the development of paralysis in MS as well as elevated glucose in diabetes

“This finding shows the potential of using a dietary l h l i disupplement to help treat autoimmune diseases. Most

importantly, we understand how this sugar‐based supplement inhibits the cells that attack the body, making metabolic therapy a rational approach to prevent or treat these debilitating diseases.”

Another recent study found improvement in 75% of children with treatment‐resistant autoimmune inflammatory bowel disease after a two year course of NAG.


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Affects viscosity of mucous membranes in yrespiratory, reproductive, and intestinal tracts.

Protects against colonization/infection bacteria in respiratory epithelial cells.p y p Mason CM J Lab Clin Med. 1992 Nov; 120: 740‐745

Researchers at the University of New Hampshire Glycomics Center led by immunologist Jeffrey Ravetch of Rockefeller University reported in the 13 May 2008 edition of the journal Science a etc o oc e e e U e s ty epo ted t e 3 ay 008 ed t o o t e jou a Sc e cethat they had identified a specific carbohydrate structure that confers anti‐inflammatory activity to a glycoprotein antibody that could lead to improved treatment of autoimmune diseases like lupus or rheumatoid arthritis.

“The work revolves around immunoglobulin G (IgG), the most abundant antibody in blood plasma. Intravenous immunoglobulin (IVIG) has trace amounts of this very active material which effectively relieves inflammatory affects of lupus, rheumatoid arthritis, asthma, and other autoimmune diseases. But because of the trace amounts of active material, effective doses of IVIG need to be very high leading to unwanted side effects.”

“This study involved rebuilding the human IVIG into a fully active molecule with a slight modification to a carbohydrate residue. These carbohydrate structures are linked to the immunoglobulin and referred to as glycans, and on the tip of this glycan is a specifically linked SialicAcid. All the Sialic Acid on IVIG was converted to the active linkage that confers anti‐inflammatory properties.”


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The Glycomics Center helped Ravetch pinpoint exactly how Sialic Acid was linked, which let the researchers engineer a synthetic human antibody that mimicked the linkages in let the researchers engineer a synthetic human antibody that mimicked the linkages in IgG, providing an IVIG with enhanced activity for treatment of autoimmune diseases. In the Science paper, the researchers report that when given to arthritic mice, the engineered IgG was about 30 times more effective than IVIG.

“Understanding and analyzing the exact structure of Sialic Acid was the contribution of the UNH Glycomics Center, headed by director and research professor Vernon Reinhold. The center has developed tools and protocols using multidimensional mass

d i h d f i l l i hi f h spectrometry to determine the structure and functional relationships of these carbohydrates. Reinhold notes that while most biopolymers are linear and thus relatively easy to sequence, bush‐shaped carbohydrates have proved challenging.”

“With sequential mass spectrometry, we systematically untangle this bush,” says Reinhold. “We take it down to the trunk then try to put it back together to determineits structure.”

Beyond this work with IgG, the Glycomics Center is demystifying the carbohydrate connections in cancer that contributes to metastatic carbohydrate connections in cancer that contributes to metastatic growth and in avian flu where Sialic residues on airway surface tissues serve as doorways for viral entry.

“Carbohydrates are the glue that pulls things together, the cell surface matrix in which cells communicate and they provide the connections for signal transduction. It’s only been within the last decade that we’ve realized that such structures are critical for all kinds of biological function,” says Reinhold. “Now that we can define precise structures, we can begin to understand their function. This structure‐functional relationship will have a huge impact on our health in respect to immune regulation.”


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Take enough (amount) Take enough (amount)

For long enough (time)

Cell apoptosis:“A time to live and a time to die”—programmed cell death

THE PATH OF DISCOVERYTHE PATH OF DISCOVERY


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“The difference in oligosaccharide chains could affect adhesive interactions between cancer cells and their normal parent tissue interactions between cancer cells and their normal parent tissue cells, contributing to metastasis.”

—Harper’s Illustrated Biochemistry (p 535)

Nutraceutical dietary supplements: Do not inhibit tumor cell destruction by radiation and chemotherapy—

actually enhances it! Protect normal cells from radiation and cytotoxic damageProtect normal cells from radiation and cytotoxic damage. Induce reductions in tumor mass in malignancies resistant to

all treatments. Improve quality of life for patients by reducing toxicity and side effects

from radiation and chemotherapy. G. Hyland, M.D., D. Miller, M.T., MedcenterOne, Dept. Radiation Oncology, Bismark,

North Dakota

National Institute of Health (NIH) Press Release, 21 Aug 2007, announced new $15.5 Million, Five‐Year Initiative to study the Glycobiology of Cancer$15.5 Million, Five Year Initiative to study the Glycobiology of Cancer

Seven Tumor Glycome Laboratories projects funded to discover, develop, and clinically validate cancer biomarkers by targeting the carbohydrate (glycan) part of a molecule.

“Scientists have long recognized that certain sugar structures, which are attached to protein and lipid molecules, may be important as markers for cancer development,” said NCI Director John E. Niederhuber

“While this area has compelling scientific interest, its biological and chemical p g , gcomplexities have often discouraged investigation. Today with the advent of advanced technologies to conduct protein and carbohydrate chemistry, research into this intriguing area has experienced renewed interest.” (Niederhuber)

Analysis may improve the value of tests such as prostate‐specific antigen (PSA), CA‐125, and carcinoembryonic antigen used in prostate, ovarian, and colon cancer detection respectively


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Seven NCI‐funded projects searching for glycan‐based biomarkers for melanoma, breast, ovarian, lung, prostate, colon, and pancreatic cancers: Discovery and clinical validation of cancer biomarkers (Margaret Huflejt, PhD, Cellexicon,

Inc, La Jolla, CA) Immunogenic sugar moieties of prostate cancers (DenongWang, M.D., PhD, Stanford

University, Palo Alto, CA) Early cancer detection and prognosis through glycomics (Milos Novotny, PhD, Indiana

University, Bloomington, IN) Glycan markers for the early detection of breast cancer (William Hancock, PhD, ScD,

Northeastern University, Boston, MA) Tumor glycomics laboratory for discovery of pancreatic cancer markers (J Michael Pierce Tumor glycomics laboratory for discovery of pancreatic cancer markers (J. Michael Pierce,

PhD, University of Georgia, Athens, GA) Autoantibodies against glycopeptide epitopes as serum biomarkers of cancer (Michael

Hollingsworth, PhD, University of Nebraska, Lincoln, NE) Neu5Gc (Sialic Acid) and AntiNeu5Gc antibodies for detection of cancer (Ajit Varki, M.D.,

University of California, San Diego, CA)

If you would like to further investigate this science or if you have any questions, y y q ,

please contact the person whoshared or recommended this class

C O P Y R I G H T © 2 0 1 0 A N G I E ’ S O P T I O N , I N C . A L L R I G H T S R E S E R V E DC O P Y R I G H T © 2 0 1 0 A N G I E ’ S O P T I O N , I N C . A L L R I G H T S R E S E R V E D

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